Method for producing fucoidan from laminaria

FIELD: bioengineering.

SUBSTANCE: method involves producing acids polysaccharide fucoidan from laminaria by crushing raw material by treating it with 0.5-1.0% food acid solution during 4-5 h and subjecting acid extract to ultrafiltration on membrane of 100-300 kDa with following end product drying by applying lipophilic or spraying or vacuum drying method.

EFFECT: high purity of end product.

 

The invention relates to the processing of brown algae for the production of biologically active polysaccharide, in particular also for use in preventive medicine, Microbiology, chemical-pharmaceutical, cosmetic, food industry, etc.

Acidic polysaccharide fucoidan is a water-soluble high-molecular compound that contains many of the side chains. He possesses various biological activities, including anticoagulation, onkoprofilakticheskim, anti-virus, lipid-lowering.

Known methods for producing from brown algae acidic polysaccharide fucoidan:

- RF patent №2247574 from 26.12.2002, IPC AC 35/80 "Means, with anticoagulant and immunotropic action, including the processing of brown seaweed Fucus evanescens ethanol, then drying and grinding, water treatment, first at room temperature and then at a temperature of 50-60°C. the Concentration of the extract, the sludge separation, obtaining sodium salt of fucoidan by planting its benchmark, washing with water and drying also ethanol;

- RF patent №2135518 from 17.06.1998 "Method of obtaining water-soluble polysaccharides of brown algae". The method provides for processing fresh or frozen, or dried seaweed solvents (ethanol, acetone, chloroform), is using serial extraction of polysaccharides 0.1 N hydrochloric acid at room temperature and water 50-60° C. Receipt of fucoidan is performed using hydrophobic chromatography;

- RF patent №2028153 from 09.02.1995 "Method of obtaining biologically active substances from kelp". The method involves the processing of fresh, frozen or dried brown algae alcohol solution followed by precipitation of the alcohol solution of the complex of laminaran with fucoidan. For the Department of fucoidan residue is treated with 0.4% solution of hydrochloric acid at room temperature, and water at a temperature of 50-60°C for 2 hours; then to the resulting solution was added 0.5 M solution of cetavlon for transferring the acidic polysaccharides in the sediment. The precipitate is dissolved in 0.4% solution of hydrochloric acid, and then treated with a solution of ethanol with getting fucoidan;

- RF patent №2240816 from 28.07.03, while brown algae treated with ethanol, processed seaweed is extracted with hydrochloric acid, the extract was concentrated to 1/5 of the original volume by ultrafiltration plant with a membrane having pores of 5 to 100 kDa, neutralized to pH 6.0 with sodium hydroxide solution and evaporated on a rotary evaporator to 1 L. the resulting concentrate is neutralized, and then ethanol precipitated fucoidan and dried;

- RF patent №2233104, 09.12.02 "Method for integrated processing of brown algae with obtaining iodine and polishuri the data products", this brown algae is extracted with a water-alcohol solution, the solid residue is treated with 1% solution of hydrochloric acid for 6-10 hours at room temperature. The resulting mass is heated to a temperature of 45-55°C for 4-6 hours Obtained acid extract is first neutralized to pH 7, evaporated under vacuum at a temperature of 50-55°and then fucoidan allocate by precipitating it with ethanol and dried.

The disadvantages of these methods is that the raw material is first treated with an alcohol-based solvent for the acidic polysaccharide fucoidan use or expensive chemical substances (alcohol or cetavlon), or expensive equipment, namely, hydrophobic chromatography, rotary evaporators. The drying is carried out mainly by alcohol.

The closest to the technical essence of the method of obtaining acidic polysaccharides is the method according to patent No. 2041656 from 23.10 1991, IPC 6 A23L 1/337 "Method of obtaining food preparation from laminaria algae, including grinding of brown algae, processing their food acid at pH 1-3 2-3 hours at room temperature, obtaining a gel-like product, which includes an acidic polysaccharide fucoidan.

The disadvantage of this method is to obtain only a comprehensive acidic polisher is Yes, composed as low molecular weight products, and high-molecular polysaccharides, in particular fucoidan.

The objective of this invention is to provide a clean acidic polysaccharide fucoidan, preserving its natural properties, as well as increasing the purity of the final product.

The problem is solved by the grinding of raw materials, processing of brown algae acid and basic solution with a pH 1-3, separating the acid extract and extract of the final product, according to the invention the processing of brown algae hold 0.5-1% solution of acid for 4-5 hours at room temperature, the final product is produced by disposable ultrafiltration membrane from 100 to 300 kDa, and then dried. Moreover, the drying is carried out using freeze drying, or spray, or vacuum.

The proposed solution meets the criteria of "novelty", "inventive step" and "industrial application".

The method consists in the following.

Seaweed raw (or dry after soaking in water at 80°C for 2-3 hours) are ground in a meat-grinder and carry out the acid treatment of 0.5-1.0% solution of edible acid (pH 1-3) for 4-5 hours at room temperature. The acid extract is separated from the solids by filtration through a sieve and tissue is iltr and sent to ultrafiltration with a membrane of 100 to 300 kDa, the final product is dried.

New the claimed technical solution is that the feedstock - brown algae treated with 0.5-1.0% solution of acid for 4-5 hours at room temperature, and the final product produced by disposable ultrafiltration membrane from 100 to 300 kDa acidic extract, followed by drying.

Processing raw materials of 0.5-1.0% acid solution with a pH of 1-3 for 4-5 hours at room temperature makes it possible to retain the native properties of the final.

The use of disposable ultrafiltration membrane from 100 to 300 kDa allows you to receive no additional cost high molecular weight acidic polysaccharide fucoidan with a molecular weight of from 100,000 to 300,000 daltons, completely get rid of low-molecular-weight polysaccharides, reducing native activity of the drug and cause side effects, including allergies and get a polysaccharide, which has high biological activity, namely anti-inflammatory, antioxidant and anticoagulant, anti-virus. The yield of the final product is 6.0-8.0% of the dry raw material.

Drying of the final product is carried out without the use of alcohol, which significantly reduces the drying time and effort.

The inventive method is characterized by simplicity and adaptability, because it includes mostly extracts the Yu and ultrafiltration. Using the last dose get native fucoidan with a purity of 92-96%.

Quantitative determination of fucoidan was performed by the spectrophotometric method by the reaction of interaction of fucoidan with thioglycolic acid in the medium of sulfuric acid and analysis of a sample of a substance by NMR nuclei13and N1.

Received fucoidan is a white powder soluble in water and are well absorbed in the gastrointestinal tract. This is due to the fact that the molecular weight of the fucoidan is 100000-300000 daltons. The molecular weight and the number of sulfate groups provides its dissolution in water and acidic solutions, which leads to the possibility of therapeutic application of fucoidan obtained by the present technology.

Acute toxicity was studied on weinbrenner mice by adding it to food and the introduction into the skin. The obtained extract is non-toxic when it is received at doses up to 500 mg/kg

Specific example

Example 1.

10 kg of raw laminaria ground in a meat-grinder and placed in an acidic solution (12 l of water and 1 l of 80% acetic acid at pH 3 to 5 hours at room temperature). The resulting mixture was filtered through a sieve and fabric filter. The obtained acidic extract is passed through an ultrafiltration installation with a membrane having pore kDa. In the process of ultrafiltration get fucoidan with a molecular weight of 200,000 daltons, which is dried using freeze drying. The final product - fucoidan is a white powder, soluble in water.

The output is70 g
The purity of the product94%.

Example 2.

10 kg of frozen raw kelp thawed, minced in a meat-grinder and placed in a 1% solution of hydrochloric acid for 4 hours at room temperature. The resulting mixture was filtered through a sieve and fabric filter, then the acidic extract is passed through an ultrafiltration installation with a membrane having pores of 100 kDa. In the process of ultrafiltration get fucoidan with a molecular weight of 100,000 daltons, which is then dried using spray drying.

The product is a white powder, soluble in water.

The output is60 g
The purity of the product- 96%.

Example 3

1.3 kg of dry kelp soaked in 15 l of water at 80°C for 2-3 hours, ground in a meat-grinder and placed in a 0.8% solution of hydrochloric acid for 5 hours at room temperature. Then received MES filtered through a sieve and fabric filter, separating the acid extract and put it through the ultrafiltration installation with a membrane having pores of 300 kDa. After ultrafiltration get fucoidan with a molecular weight of 300,000 daltons, which is then dried using a drum drying.

The product is a white powder, soluble in water.

The output is80 g
The purity of the product92%.

Currently experienced on the basis of FSUE "TINRO-Center" the pilot batch of fucoidan in the amount of 150,

1. The method of obtaining of fucoidan from laminaria, including the grinding of raw materials, processing the acid solution, separating the acid extract and obtain from it the final product, characterized in that the processing of kelp conduct of 0.5-1.0%solution of edible acid for 4-5 h, and the final product obtained by ultrafiltration of acid extract on membrane with pore size of 100 to 300 kDa, and then dried.

2. The method according to claim 1, characterized in that the drying of the final product is carried out either freeze or spray or vacuum drying.



 

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